scholarly journals Inhibition of henipavirus infection by Nipah virus attachment glycoprotein occurs without cell-surface downregulation of ephrin-B2 or ephrin-B3

2007 ◽  
Vol 88 (2) ◽  
pp. 582-591 ◽  
Author(s):  
Bevan Sawatsky ◽  
Allen Grolla ◽  
Nina Kuzenko ◽  
Hana Weingartl ◽  
Markus Czub
Keyword(s):  
Cell Surface ◽  
Nipah Virus ◽  
Functional Expression ◽  
Hendra Virus ◽  
Pcr Analysis ◽  
Protein G ◽  
Attachment Protein ◽  
Genetic Characteristics ◽  
Virus Attachment ◽  
Fusion Inhibition

Nipah virus (NiV) and Hendra virus (HeV) are newly identified members of the family Paramyxoviridae and have been classified in the new genus Henipavirus based on unique genetic characteristics distinct from other paramyxoviruses. Transgenic cell lines were generated that expressed either the attachment protein (G) or the fusion protein (F) of NiV. Functional expression of NiV F and G was verified by complementation with the corresponding glycoprotein, which resulted in the development of syncytia. When exposed to NiV and HeV, expression of NiV G in Crandall feline kidney cells resulted in a qualitative inhibition of both cytopathic effect (CPE) and cell death by both viruses. RT-PCR analysis of surviving exposed cells showed a complete absence of viral positive-sense mRNA and genomic negative-sense viral RNA. Cells expressing NiV G were also unable to fuse with cells co-expressing NiV F and G in a fluorescent fusion inhibition assay. Cell-surface staining for the cellular receptors for NiV and HeV (ephrin-B2 and ephrin-B3) indicated that they were located on the surface of cells, regardless of NiV G expression or infection by NiV. These results indicated that viral interference can be established for henipaviruses and requires only the expression of the attachment protein, G. Furthermore, it was found that this interference probably occurs at the level of virus entry, as fusion was not observed in cells expressing NiV G. Finally, expression of NiV G by either transient transfection or NiV infection did not alter the cell-surface levels of the two known viral receptors.

scholarly journals Location of, immunogenicity of and relationships between neutralization epitopes on the attachment protein (G) of Hendra virus

2005 ◽  
Vol 86 (10) ◽  
pp. 2839-2848 ◽  
Author(s):  
John R. White ◽  
Victoria Boyd ◽  
Gary S. Crameri ◽  
Christine J. Duch ◽  
Ryan K. van Laar ◽  
...  
Keyword(s):  
Amino Acid ◽  
Measles Virus ◽  
Structural Model ◽  
Nipah Virus ◽  
Hendra Virus ◽  
Protein G ◽  
Head Region ◽  
Attachment Protein ◽  
Virus Attachment ◽  
Discontinuous Epitope

Epitopes involved in a protective immune response to Hendra virus (HeV) (Henipavirus, Paramxyoviridae) were investigated by generating five neutralizing monoclonal antibodies (mAbs) to the virus attachment protein (G) of HeV (HeV G) and sequencing of the G gene of groups of neutralization-escape variants selected with each mAb. Amino acid substitutions occurred at eight distinct sites on HeV G. Relationships between these sites were investigated in binding and neutralization assays using heterologous combinations of variants and mAbs. The sites were also mapped to a proposed structural model for the attachment proteins of Paramyxoviridae. Their specific locations and the nature of their interactions with the mAb panel provided the first functional evidence that HeV G in fact resembled the proposed structure. Four sites (aa 183–185, 417, 447 and 570) contributed to a major discontinuous epitope, on the base of the globular head, that was similar to immunodominant virus neutralization sites found in other paramyxoviruses. Amino acid similarity between HeV and Nipah virus was relatively highly conserved at these sites but decreased significantly at the other sites identified in this study. These included another discontinuous epitope on the base of the head region defined by sites aa 289 and 324 and well separated epitopes on the top of the head at sites aa 191–195 and 385–356. The latter epitope corresponded to immunodominant neutralization sites found in Rinderpest virus and Measles virus.

scholarly journals Feline Model of Acute Nipah Virus Infection and Protection with a Soluble Glycoprotein-Based Subunit Vaccine

2006 ◽  
Vol 80 (24) ◽  
pp. 12293-12302 ◽  
Author(s):  
Bruce A. Mungall ◽  
Deborah Middleton ◽  
Gary Crameri ◽  
John Bingham ◽  
Kim Halpin ◽  
...  
Keyword(s):  
Subunit Vaccine ◽  
Case Reports ◽  
Mammalian Species ◽  
Nipah Virus ◽  
Disease Onset ◽  
Hendra Virus ◽  
Clinical Disease ◽  
Pcr Analysis ◽  
Wide Range ◽  
Feline Model

ABSTRACT Nipah virus (NiV) and Hendra virus (HeV) are paramyxoviruses capable of causing considerable morbidity and mortality in a number of mammalian species, including humans. Case reports from outbreaks and previous challenge experiments have suggested that cats were highly susceptible to NiV infection, responding with a severe respiratory disease and systemic infection. Here we have assessed the cat as a model of experimental NiV infection and use it in the evaluation of a subunit vaccine comprised of soluble G glycoprotein (sG). Two groups of two adult cats each were inoculated subcutaneously with either 500 or 5,000 50% tissue culture infective dose(s) (TCID50) of NiV. Animals were monitored closely for disease onset, and extensive analysis was conducted on samples and tissues taken during infection and at necropsy to determine viral load and tissue tropism. All animals developed clinical disease 6 to 9 days postinfection, a finding consistent with previous observations. In a subsequent experiment, two cats were immunized with HeV sG and two were immunized with NiV sG. Homologous serum neutralizing titers were greater than 1:20,000, and heterologous titers were greater than 1:20,000 to 16-fold lower. Immunized animals and two additional naive controls were then challenged subcutaneously with 500 TCID50 of NiV. Naive animals developed clinical disease 6 to 13 days postinfection, whereas none of the immunized animals showed any sign of disease. TaqMan PCR analysis of samples from naive animals revealed considerable levels of NiV genome in a wide range of tissues, whereas the genome was evident in only two immunized cats in only four samples and well below the limit of accurate detection. These results indicate that the cat provides a consistent model for acute NiV infection and associated pathogenesis and an effective subunit vaccine strategy appears achievable.

scholarly journals Differential Rates of Protein Folding and Cellular Trafficking for the Hendra Virus F and G Proteins: Implications for F-G Complex Formation

2009 ◽  
Vol 83 (17) ◽  
pp. 8998-9001 ◽  
Author(s):  
Shannon D. Whitman ◽  
Everett Clinton Smith ◽  
Rebecca Ellis Dutch
Keyword(s):  
Endoplasmic Reticulum ◽  
Complex Formation ◽  
G Proteins ◽  
Secretory Pathway ◽  
Viral Proteins ◽  
Hendra Virus ◽  
Protein G ◽  
Cellular Transport ◽  
Attachment Protein ◽  
Viral Attachment

ABSTRACT Hendra virus F protein-promoted membrane fusion requires the presence of the viral attachment protein, G. However, events leading to the association of these glycoproteins remain unclear. Results presented here demonstrate that Hendra virus G undergoes slower secretory pathway trafficking than is observed for Hendra virus F. This slowed trafficking is not dependent on the G protein cytoplasmic tail, the presence of the G receptor ephrin B2, or interaction with other viral proteins. Instead, Hendra virus G was found to undergo intrinsically slow oligomerization within the endoplasmic reticulum. These results suggest that the critical F-G interactions occur only after the initial steps of synthesis and cellular transport.

scholarly journals Crystal Structure and Carbohydrate Analysis of Nipah Virus Attachment Glycoprotein: a Template for Antiviral and Vaccine Design

2008 ◽  
Vol 82 (23) ◽  
pp. 11628-11636 ◽  
Author(s):  
Thomas A. Bowden ◽  
Max Crispin ◽  
David J. Harvey ◽  
A. Radu Aricescu ◽  
Jonathan M. Grimes ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Ebola Virus ◽  
Nipah Virus ◽  
Glycosylation Site ◽  
Hendra Virus ◽  
Molecular Surface ◽  
Spectrometric Analysis ◽  
Viral Attachment ◽  
Virus Attachment ◽  
Host Interaction

ABSTRACT Two members of the paramyxovirus family, Nipah virus (NiV) and Hendra virus (HeV), are recent additions to a growing number of agents of emergent diseases which use bats as a natural host. Identification of ephrin-B2 and ephrin-B3 as cellular receptors for these viruses has enabled the development of immunotherapeutic reagents which prevent virus attachment and subsequent fusion. Here we present the structural analysis of the protein and carbohydrate components of the unbound viral attachment glycoprotein of NiV glycoprotein (NiV-G) at a 2.2-Å resolution. Comparison with its ephrin-B2-bound form reveals that conformational changes within the envelope glycoprotein are required to achieve viral attachment. Structural differences are particularly pronounced in the 579-590 loop, a major component of the ephrin binding surface. In addition, the 236-245 loop is rather disordered in the unbound structure. We extend our structural characterization of NiV-G with mass spectrometric analysis of the carbohydrate moieties. We demonstrate that NiV-G is largely devoid of the oligomannose-type glycans that in viruses such as human immunodeficiency virus type 1 and Ebola virus influence viral tropism and the host immune response. Nevertheless, we find putative ligands for the endothelial cell lectin, LSECtin. Finally, by mapping structural conservation and glycosylation site positions from other members of the paramyxovirus family, we suggest the molecular surface involved in oligomerization. These results suggest possible pathways of virus-host interaction and strategies for the optimization of recombinant vaccines.

scholarly journals Single Amino Acid Changes in the Nipah and Hendra Virus Attachment Glycoproteins Distinguish EphrinB2 from EphrinB3 Usage

2007 ◽  
Vol 81 (19) ◽  
pp. 10804-10814 ◽  
Author(s):  
Oscar A. Negrete ◽  
David Chu ◽  
Hector C. Aguilar ◽  
Benhur Lee
Keyword(s):  
Brain Stem ◽  
Viral Entry ◽  
Nipah Virus ◽  
Chimeric Protein ◽  
Hendra Virus ◽  
Single Amino Acid ◽  
Virus Attachment ◽  
Receptor Usage ◽  
Binding Determinants ◽  
The Brain

ABSTRACT The henipaviruses, Nipah virus (NiV) and Hendra virus (HeV), are lethal emerging paramyxoviruses. EphrinB2 and ephrinB3 have been identified as receptors for henipavirus entry. NiV and HeV share similar cellular tropisms and likely use an identical receptor set, although a quantitative comparison of receptor usage by NiV and HeV has not been reported. Here we show that (i) soluble NiV attachment protein G (sNiV-G) bound to cell surface-expressed ephrinB3 with a 30-fold higher affinity than that of sHeV-G, (ii) NiV envelope pseudotyped reporter virus (NiVpp) entered ephrinB3-expressing cells much more efficiently than did HeV pseudotyped particles (HeVpp), and (iii) NiVpp but not HeVpp entry was inhibited efficiently by soluble ephrinB3. These data underscore the finding that NiV uses ephrinB3 more efficiently than does HeV. Henipavirus G chimeric protein analysis implicated residue 507 in the G ectodomain in efficient ephrinB3 usage. Curiously, alternative versions of published HeV-G sequences show variations at residue 507 that can clearly affect ephrinB3 but not ephrinB2 usage. We further defined surrounding mutations (W504A and E505A) that diminished ephrinB3-dependent binding and viral entry without compromising ephrinB2 receptor usage and another mutation (E533Q) that abrogated both ephrinB2 and -B3 usage. Our results suggest that ephrinB2 and -B3 binding determinants on henipavirus G are distinct and dissociable. Global expression analysis showed that ephrinB3, but not ephrinB2, is expressed in the brain stem. Thus, ephrinB3-mediated viral entry and pathology may underlie the severe brain stem neuronal dysfunction seen in fatal Nipah viral encephalitis. Characterizing the determinants of ephrinB2 versus -B3 usage will further our understanding of henipavirus pathogenesis.

Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2

2008 ◽  
Vol 15 (6) ◽  
pp. 567-572 ◽  
Author(s):  
Thomas A Bowden ◽  
A Radu Aricescu ◽  
Robert J C Gilbert ◽  
Jonathan M Grimes ◽  
E Yvonne Jones ◽  
...  
Keyword(s):  
Cell Surface ◽  
Hendra Virus ◽  
Cell Surface Receptor ◽  
Structural Basis ◽  
Virus Attachment ◽  
Surface Receptor

scholarly journals A single dose investigational subunit vaccine for human use against Nipah virus and Hendra virus

npj Vaccines ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Thomas W. Geisbert ◽  
Kathryn Bobb ◽  
Viktoriya Borisevich ◽  
Joan B. Geisbert ◽  
Krystle N. Agans ◽  
...  
Keyword(s):  
Single Dose ◽  
Subunit Vaccine ◽  
Nipah Virus ◽  
Hendra Virus ◽  
Vaccine Formulation ◽  
Protein Subunit ◽  
Primate Model ◽  
Virus Attachment ◽  
A Subunit ◽  
Human Vaccine

AbstractNipah and Hendra viruses are highly pathogenic bat-borne paramyxoviruses recently included in the WHO Blueprint priority diseases list. A fully registered horse anti-Hendra virus subunit vaccine has been in use in Australia since 2012. Based on the same immunogen, the Hendra virus attachment glycoprotein ectodomain, a subunit vaccine formulation for use in people is now in a Phase I clinical trial. We report that a single dose vaccination regimen of this human vaccine formulation protects against otherwise lethal challenges of either Hendra or Nipah virus in a nonhuman primate model. The protection against the Nipah Bangladesh strain begins as soon as 7 days post immunization with low dose of 0.1 mg protein subunit. Our data suggest this human vaccine could be utilized as efficient emergency vaccine to disrupt potential spreading of Nipah disease in an outbreak setting.

scholarly journals Expression, characterisation and antigenicity of a truncated Hendra virus attachment protein expressed in the protozoan host Leishmania tarentolae

2016 ◽  
Vol 228 ◽  
pp. 48-54 ◽  
Author(s):  
Kerstin Fischer ◽  
Vinicius Pinho dos Reis ◽  
Stefan Finke ◽  
Lucie Sauerhering ◽  
Eileen Stroh ◽  
...  
Keyword(s):  
Hendra Virus ◽  
Attachment Protein ◽  
Leishmania Tarentolae ◽  
Virus Attachment

scholarly journals Characterization of African bat henipavirus GH-M74a glycoproteins

2014 ◽  
Vol 95 (3) ◽  
pp. 539-548 ◽  
Author(s):  
Michael Weis ◽  
Laura Behner ◽  
Markus Hoffmann ◽  
Nadine Krüger ◽  
Georg Herrler ◽  
...  
Keyword(s):  
G Protein ◽  
Mammalian Cells ◽  
Nipah Virus ◽  
Syncytium Formation ◽  
Cell Types ◽  
Functional Expression ◽  
Surface Expression ◽  
Hendra Virus ◽  
Proteolytic Activation ◽  
F Protein

In recent years, novel henipavirus-related sequences have been identified in bats in Africa. To evaluate the potential of African bat henipaviruses to spread in non-bat mammalian cells, we compared the biological functions of the surface glycoproteins G and F of the prototype African henipavirus GH-M74a with those of the glycoproteins of Nipah virus (NiV), a well-characterized pathogenic member of the henipavirus genus. Glycoproteins are central determinants for virus tropism, as efficient binding of henipavirus G proteins to cellular ephrin receptors and functional expression of fusion-competent F proteins are indispensable prerequisites for virus entry and cell-to-cell spread. In this study, we analysed the ability of the GH-M74a G and F proteins to cause cell-to-cell fusion in mammalian cell types readily permissive to NiV or Hendra virus infections. Except for limited syncytium formation in a bat cell line derived from Hypsignathus monstrosus, HypNi/1.1 cells, we did not observe any fusion. The highly restricted fusion activity was predominantly due to the F protein. Whilst GH-M74a G protein was found to interact with the main henipavirus receptor ephrin-B2 and induced syncytia upon co-expression with heterotypic NiV F protein, GH-M74a F protein did not cause evident fusion in the presence of heterotypic NiV G protein. Pulse–chase and surface biotinylation analyses revealed delayed F cleavage kinetics with a reduced expression of cleaved and fusion-active GH-M74a F protein on the cell surface. Thus, the F protein of GH-M74a showed a functional defect that is most likely caused by impaired trafficking leading to less efficient proteolytic activation and surface expression.

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